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Journal of Forensic Medicine ; (6): 123-125, 2012.
Article Dans Chinois | WPRIM | ID: wpr-983726

Résumé

OBJECTIVE@#To establish optimal amplification conditions with the application of 0.2 mL test tube in single cell separation and inspection.@*METHODS@#Oral epithelium cell suspension was prepared. Five or ten cells were collected with 0.2 mL test tube. Then DNA were amplified with Identifiler Plus kit in three different conditions in which the proteinase K addition, gold enzyme concentration in PCR reaction, and PCR reaction cycles were adjusted separately. Finally the detection rate, allelic dropout rate and artificial alleles were compared among the groups.@*RESULTS@#In these 3 different conditions: addition of proteinase K, addition of 0.4 microL gold enzyme in PCR reaction, and use of 32 cycles, the detection rate was higher and allelic dropout rate was lower than the other conditions.@*CONCLUSION@#In single cell separation and inspection, the usage of 0.2 mL test tube could be an effective supplement to chip-low volume amplification.


Sujets)
Humains , Allèles , Séparation cellulaire/méthodes , ADN/génétique , Profilage d'ADN/méthodes , Endopeptidase K/administration et posologie , Enzymes/administration et posologie , Cellules épithéliales/cytologie , Études de faisabilité , Génétique légale , Génotype , Muqueuse de la bouche/cytologie , Techniques d'amplification d'acides nucléiques , Réaction de polymérisation en chaîne/méthodes , Sensibilité et spécificité
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